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Plant Physiol, May 2002, Vol. 129, pp. 13-22
Agrobacterium tumefaciens-Mediated Transformation of
Maize Embryos Using a Standard Binary Vector
System1
Bronwyn R.
Frame,
Huixia
Shou,
Rachel K.
Chikwamba,
Zhanyuan
Zhang,2
Chengbin
Xiang,
Tina M.
Fonger,
Sue Ellen K.
Pegg,3
Baochun
Li,4
Dan S.
Nettleton,
Deqing
Pei, and
Kan
Wang*
Plant Transformation Facility, Departments of Agronomy (B.R.F.,
H.S., R.K.C., Z.Z., T.M.F., S.E.K.P., B.L., K.W.), Botany (C.X.), and
Statistics (D.S.N., D.P.), Iowa State University, Ames, Iowa
50011
We have achieved routine transformation of maize (Zea
mays) using an Agrobacterium tumefaciens
standard binary (non-super binary) vector system. Immature zygotic
embryos of the hybrid line Hi II were infected with A.
tumefaciens strain EHA101 harboring a standard binary vector
and cocultivated in the presence of 400 mg L 1
L-cysteine. Inclusion of L-cysteine in
cocultivation medium lead to an improvement in transient
 -glucuronidase expression observed in targeted cells and a
significant increase in stable transformation efficiency, but was
associated with a decrease in embryo response after cocultivation. The
average stable transformation efficiency (no. of bialaphos-resistant
events recovered per 100 embryos infected) of the present protocol was
5.5%. Southern-blot and progeny analyses confirmed the integration,
expression, and inheritance of the bar and
gus transgenes in R0, R1, and
R2 generations of transgenic events. To our knowledge, this
represents the first report in which fertile, stable transgenic maize
has been routinely produced using an A. tumefaciens
standard binary vector system.
1
This work was supported by the Iowa Corn
Promotion Board; by the Agricultural Experiment Station, the Office of
Biotechnology, and the Plant Science Institute of Iowa State
University; by the Baker Endowment Advisory Council for Excellence in
Agronomy; by the National Science Foundation (grant no. DBI-0077692 to
H.S. and B.L.); and by the Rockefeller Foundation (to R.K.C.). This is
journal paper no. J-19539 of the Iowa Agriculture and Home Economics
Experiment Station (Ames), project no. 3703, supported by the Hatch Act
and the State of Iowa.
2
Present address: Plant Transformation Facility,
University of Missouri-Columbia, Columbia, MO 65211.
3
Present address: Department of Agriculture, Western
Illinois University, 1 University Circle, Macomb, IL 61455.
4
Present address: Tobacco and Health Research Institute,
University of Kentucky, Cooper and University Drives, Lexington, KY 40546.
*
Corresponding author; e-mail kanwang{at}iastate.edu; fax
515-294-2299.
© 2002 American Society of Plant Physiologists
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